ISSR(inter simple sequence repeat) markers were used to detected genetic diversity among wheat cultivars(or breeding lines) of Huang-Huai Winter Wheat Region of China.Totally 96 alleles of 11 ISSR markers were detected among 96 cultivars with the average of 8.7 alleles per ISSR marker,and the number of alleles per primer ranged from 3 to 23.The value of allelic polymorphism information content(PIC) ranged from 0.601 to 0.941,on the average of 0.791 per primer.These results showed that ISSR is a suitable molecular technique for genetic diversity analysis of wheat germplasm resources.The value of Nei's genetic similarity(GS) indexes of 96 cultivars based on the ISSR data varied from 0.35 to 0.91,with an average of 0.60,indicating that the genetic diversity among cultivars of Huang-Huai Wheat Region is abundant.Cluster analysis showed that the cultivars tested in this study could be clustered into eight groups and fourteen sub-groups,which generally do not agree with pedigree relations and regions of origin.